grothlab/crepas

a reproducible nascent chromatin sequencing analysis pipeline for epigenome replication studies.

https://github.com/grothlab/crepas/tree/mainView on GitHub

Rendered from nextflow_schema.json. Parameters are passed on the command line as --name value.

Define where the pipeline should find input data and save output data.

Path to comma-separated file containing information about the samples in the experiment.
required
type: string
pattern: ^\S+\.(csv|tsv|yaml|json)$
Help text

You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. See usage docs.

Estimated fragment size used to extend single-end reads.
type: integer
default: 250
Platform/technology used to produce the reads. Corresponds to the PL tag in the SAM/BAM file header.
type: string
Help text

See the SAM format specification. Valid values: CAPILLARY, DNBSEQ (MGI/BGI), ELEMENT, HELICOS, ILLUMINA, IONTORRENT, LS454, ONT (Oxford Nanopore), PACBIO (Pacific Biosciences), SINGULAR, SOLID, and ULTIMA. This field should be omitted when the technology is not in this list (though the PM field may still be present in this case) or is unknown.

Sequencing center information to be added to read group of BAM files.
type: string
Read length used to calculate MACS3 genome size for peak calling if --macs_gsize isn't provided.
type: integer
options: 50, 75, 100, 150, 200
default: 75
The output directory where the results will be saved. You have to use absolute paths to storage on Cloud infrastructure.
required
type: string
Email address for completion summary.
type: string
pattern: ^([a-zA-Z0-9_\-\.]+)@([a-zA-Z0-9_\-\.]+)\.([a-zA-Z]{2,5})$
Help text

Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (~/.nextflow/config) then you don't need to specify this on the command line for every run.

MultiQC report title. Printed as page header, used for filename if not otherwise specified.
type: string

Reference genome related files and options required for the workflow.

Name of reference genome (e.g., GRCh38, hg19, mm10).
required
type: string
Path to FASTA genome file. Required unless --hybrid_fasta is provided or the file is supplied by a reference genome config (iGenomes/Refgenie).
type: string
pattern: ^\S+\.fn?a(sta)?(\.gz)?$
Help text

If you don't have the appropriate alignment index available this will be generated for you automatically based on this file. Combine with --save_reference to save alignment index for future runs.

Path to exogenous (spike-in) FASTA genome file. If you provide this, the a hybrid fasta reference file will be generated and used for alignment index building.
type: string
pattern: ^\S+\.fn?a(sta)?(\.gz)?$
Path to a pre-built hybrid (endogenous + spike-in) FASTA genome file, with spike-in contigs named <chromosome_name>_<spikein_genome>. If provided, this is used directly instead of generating a hybrid fasta from --fasta/--spikein_fasta. Mutually exclusive with --fasta: providing both raises an error.
type: string
pattern: ^\S+\.fn?a(sta)?(\.gz)?$
Path to GTF annotation file.
type: string
pattern: ^\S+\.gtf(\.gz)?$
Path to GFF3 annotation file.
type: string
pattern: ^\S+\.gff3?(\.gz)?$
Path to directory or tar.gz archive for pre-built BWA index.
type: string
Path to directory or tar.gz archive for pre-built BWAMEM2 index.
type: string
Path to directory or tar.gz archive for pre-built minibwa index.
type: string
Path to directory or tar.gz archive for pre-built Bowtie index.
type: string
Path to directory or tar.gz archive for pre-built Bowtie2 index.
type: string
Path to directory or tar.gz archive for pre-built Chromap index.
type: string
Path to directory or tar.gz archive for pre-built STAR index.
type: string
Path to directory or tar.gz archive for pre-built HISAT2 index.
type: string
Path to directory or tar.gz archive for pre-built Minimap2 index.
type: string
Path to Strobealign index file with extension '.sti'. This will be created from the FASTA file if not specified and the aligner is set to strobealign.
type: string
pattern: ^\S+\.sti(\.gz)?$
Path to BED file containing gene intervals. This will be created from the GTF file if not specified.
type: string
pattern: ^\S+\.bed(\.gz)?$
Path to BED file containing initiation zones for SCAR-seq analysis.
type: string
Path to GZIP-compressed OK-seq RFD text file. The path cannot contain spaces and must have extension '.txt.gz', '.tsv.gz' or '.bed.gz'
type: string
pattern: ^\S+\.(txt|tsv|bed)(\.gz)?$
Effective genome size parameter required by MACS3.
type: number
Help text

Effective genome size parameter required by MACS3. Provide the number written out in full (e.g. --macs_gsize 2700000000), not in scientific notation: 2.7e9.

Path to blacklist regions in BED format, used for filtering alignments.
type: string
Help text

If provided, alignments that overlap with the regions in this file will be filtered out (see ENCODE blacklists). The file should be in BED format.

Path to BED file with sparse segments for Consenrich.
type: string
Help text

Defines the annotation file of 'sparse segments' (genomic regions) from the complement of an 'inflated' BED file of known active regions. See https://github.com/nolan-h-hamilton/Consenrich/blob/5e7f287d81ac4a9bc098ae29e825a5ded07af3b8/consenrich/consenrich.py#L58C54-L58C174

Path to BED file with active regions for Consenrich.
type: string
Help text

Defines the annotation file of 'active regions' (genomic regions) for Consenrich. See https://github.com/nolan-h-hamilton/Consenrich/blob/5e7f287d81ac4a9bc098ae29e825a5ded07af3b8/consenrich/consenrich.py#L67

Path to ROCCO parameters file.
type: string
Help text

CSV file containing chromosome-specific optimization parameters. Each supported genome has a custom --params file packaged with ROCCO, but a custom file has to be manually passed to ROCCO within this pipeline. Consider using this argument to set chromosome-specific budget and gamma values custom to your data and preferences. See the ROCCO documentation for more details.

Path to splice sites file for HISAT2.
type: string
Help text

If using HISAT2 as the aligner, you can provide a list of known splice sites, which HISAT2 makes use of to align reads with small anchors. If this file is not provided, it will be generated from the GTF file.

If generated by the pipeline save the BWA index in the results directory.
type: boolean
default: true
Help text

If the BWA index is generated by the pipeline use this parameter to save it to your results folder. These can then be used for future pipeline runs, reducing processing times.

Skip indexing the GTF file with tabix before processing.
type: boolean
Help text

Set this parameter to true to skip indexing the GTF file with tabix. This may be useful if your GTF file is already indexed or if you encounter issues with tabix.

Directory / URL base for iGenomes references.
hidden
type: string
default: s3://ngi-igenomes/igenomes/
Do not load the iGenomes reference config.
hidden
type: boolean
default: true
Help text

Do not load igenomes.config when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in igenomes.config.

Whether to ignore the Refgenie reference genomes.
hidden
type: boolean
default: true
Help text

Do not use the Refgenie reference genomes, even if loaded before through config files.

Options to adjust UMI extraction and deduplication criteria.

Skip the MarkDuplicates step with Picard for samples without UMIs.
type: boolean
UMI pattern to use.
type: string
options: string, regex
default: string
Help text

More details can be found in the UMI-tools documentation.

The UMI barcode pattern to use e.g. 'NNNNNN' indicates that the first 6 nucleotides of the read are from the UMI.
type: string
default: NNNNNNNNN
Help text

More details can be found in the UMI-tools documentation.

The UMI barcode pattern to use if the UMI is located in read 2.
type: string
default: NNNNNNNNN
Separator used to separate the UMI from the read name.
type: string
default: _
Method to use to determine read groups by subsuming those with similar UMIs. All methods start by identifying the reads with the same mapping position, but treat similar yet nonidentical UMIs differently.
type: string
options: unique, percentile, directional, adjacency, cluster
default: percentile
After UMI barcode extraction discard either R1 or R2 by setting this parameter to 1 or 2, respectively.
type: integer
Get deduplication statistics.
type: boolean
default: true
UMI deduplication tool to use.
type: string
options: umicollapse, umitools
default: umitools
Save intermediate files from the UMI deduplication step.
type: boolean
Skip splitting the BAM file by chromosome for UMI deduplication.
type: boolean
Help text

BAMs are splitted by chromosome, deduplicated, and then merged back to improve parallelization of the deduplication step. Set this parameter to true to skip this splitting step. This may be useful if you encounter issues with the splitting or merging steps or if your BAM files are small and you don't think that parallelization will speed up the deduplication step.

Options to adjust adapter trimming criteria.

Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads).
type: integer
Instructs Trim Galore to remove bp from the 5' end of read 2 (paired-end reads only).
type: integer
Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed.
type: integer
Instructs Trim Galore to remove bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed.
type: integer
Instructs Trim Galore to search for the full length (33 bp) TruSeq (ChIP-seq, ChOR-seq, SCAR-seq) or Nextera (ATAC-seq) adapter sequences for both complement and reverse complements.
type: boolean
default: true
Help text

The default (false) setting will mean that the auto-detect function of Trim Galore will be used. See https://github.com/FelixKrueger/TrimGalore/blob/master/Docs/Trim_Galore_User_Guide.md#adapter-auto-detection

Instructs Trim Galore to apply the --nextseq=X option, to trim based on quality after removing poly-G tails.
type: integer
default: 20
Help text

This enables the Cutadapt --nextseq-trim=3'CUTOFF option via Trim Galore, which will set a quality cutoff (that is normally given with -q instead), but qualities of G bases are ignored. This trimming is in common for the NextSeq- and NovaSeq-platforms, where basecalls without any signal are called as high-quality G bases. Read the Cutadapt documentation for more details.

Instructs Trim Galore to use this stringency (minimum overlap) when searching for adapter sequences.
type: integer
default: 3
Help text

In other words, to reduce the number of falsely trimmed bases, the alignment algorithm in Trim Galore requires that at least this number of bases of the adapter are aligned to the read. This parameter corresponds to the --stringency option in Trim Galore and the --overlap option in Cutadapt.

Instructs Trim Galore to use this error rate when trimming.
type: number
default: 0.2
Instructs Trim Galore to use this quality cutoff when trimming.
type: integer
Instructs Trim Galore to discard reads that became shorter than this length after adapter or quality trimming. A value of 0 effectively disables this behaviour.
type: integer
default: 5
Instructs Trim Galore to discard reads that are longer than this length after adapter or quality trimming.
type: integer
Skip the adapter trimming step.
type: boolean
Help text

Use this if your input FastQ files have already been trimmed outside of the workflow or if you're very confident that there is no adapter contamination in your data.

Save the trimmed FastQ files in the results directory.
type: boolean
Help text

By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete.

Minimum number of reads required after trimming.
type: integer
default: 1

Options to adjust hard trimming criteria.

Instructs Trim Galore to hard-trim (shorten) reads to this length from their 3' end. This is performed after adapter/quality trimming.
type: integer
Instructs Trim Galore to hard-trim (shorten) reads to this length from their 5' end. This is performed after adapter/quality trimming.
type: integer
Whether to save the hard-trimmed reads (FASTQs) in the output directory.
type: boolean
default: true

Options to adjust parameters and filtering criteria for read alignments.

Specifies the alignment algorithm to use.
type: string
options: bwa, bwamem2, minibwa, bowtie, bowtie2, chromap, star, hisat2, strobealign, minimap2
default: chromap
Don't output BWA MEM alignments with score lower than this parameter.
type: integer
Don't output BWAMEM2 alignments with score lower than this parameter.
type: integer
Save the intermediate BAM files from the alignment step.
type: boolean
Help text

By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set this parameter to also save other intermediate BAM files.

Where possible, save unaligned reads from either STAR or HISAT2 to the results directory.
type: boolean
Help text

This may either be in the form of FastQ or BAM files depending on the options available for that particular tool.

Number of multimappers to map.
type: integer
Wheter to perform end-to-end alignment (only relevant for Bowtie2 and STAR). If set to false, local alignment will be performed.
type: boolean
default: true
Preset to use for minimap2 alignment. See the minimap2 documentation for more details.
type: string
options: sr
default: sr
K-mer length to use for minimap2 indexing and alignment. See the minimap2 documentation for more details.
type: integer
default: 21
Minimizer window size to use for minimap2 indexing and alignment. See the minimap2 documentation for more details.
type: integer
default: 11
Preset to use for minibwa alignment. See the minibwa documentation for more details.
type: string
options: sr, lr, adap
default: sr
Minimum seed length to use for minibwa alignment. See the minibwa documentation for more details.
type: integer
default: 19
Bandwidth to use for minibwa alignment. See the minibwa documentation for more details.
type: integer
default: 100

Options to adjust spike-in splitting criteria.

Name of the spike-in genome (if used in the experiment). The spike-in (exogenous) chromosome names in the --fasta and <aligner>_index files should have the format <chromosome_name>_<spikein_genome>, e.g. chr1_dm6.
type: string
Save intermediate files during spike-in splitting and filtering of BAMs. These include the split and filtered BAM files before coordinate-sorting.
type: boolean

Options to adjust multimapper allocation criteria.

Method to allocate multimappers. Not setting this parameter means that multimappers will not be allocated (the default).
type: string
options: allo, chromap, mmr
When using Allo for multimapper allocation, use the CNN trained on histone ChIP-seq datasets with mixed peaks, otherwise use the default (narrow). See allo for more details.
type: boolean
default: true
Save the intermediate BAM files from the multimapper allocation step.
type: boolean

Options to adjust alignment shifting criteria.

Whether to save the intermediate files from the alignment shifting (only done on ATAC-seq data).
type: boolean

Options to adjust filtering criteria for BAM files.

Whether to skip the flT3 filtering step (filtering by mapping quality, which also removes multimapping reads in most cases).
type: boolean
Whether to skip the blacklist filtering step. This serves to skip the removal of blacklisted regions from the BAM files even if a blacklist BED file is provided.
type: boolean

Options to adjust coverage and normalization criteria.

Whether to skip the SRPM normalization step. See the output documentation for more details.
type: boolean
Whether to skip the CISRPM normalization step. See the output documentation for more details.
type: boolean
This parameter allows the extension of reads to fragment size. If set, each read is extended, without exception.
type: integer
default: 250
Help text

Single-end: Requires a user specified value for the final fragment length. Reads that already exceed this fragment length will not be extended. Paired-end: Reads with mates are always extended to match the fragment size defined by the two read mates. Unmated reads, mate reads that map too far apart (>4x fragment length) or even map to different chromosomes are treated like single-end reads. The input of a fragment length value is optional. If no value is specified, it is estimated from the data (mean of the fragment size of all mate reads). Read the deepTools documentation for more details.

Size of the bins, in bases, for the output of the coverage calculation.
type: integer
default: 250
This parameter determines if non-covered regions (regions without overlapping reads) should be skipped. The default is to treat those regions as having a value of zero. See the deepTools documentation for more details.
type: boolean
Whether to skip the signal vs input comparison step.
type: boolean
Operation to perform when comparing IP and input coverage. soi is basically the same as ratio, although it is computed with a special script. For the rest of the operations, see the deepTools documentation for more details.
type: string
options: soi, log2, ratio, subtract, add, mean, reciprocal_ratio, first, second
default: soi
Minimum signal in the IP bin to be included in the signal over input (SOI) calculation. Otherwise, the the score of that bin is set to NaN.
type: number
default: 0
Minimum input signal in the input bin to be included in the signal over input (SOI) calculation. Otherwise, the the score of that bin is set to NaN.
type: number
default: 0.001
A small number to avoid x/0. Only useful together with --operation log2 or --operation ratio. See the deepTools documentation for more details.
type: number
default: 0
This parameter determines if regions with zero coverage in both the IP and input should be skipped when comparing IP and input coverage. This is determined BEFORE any applicable pseudocount is added. See the deepTools documentation for more details.
type: boolean
default: true
Comma-separated list of histone marks for which downsampling or normalization factors should be calculated using the total number of reads before the quality filtering in flT3. The total mapped reads after flT1 will be used if flT2 was not performed (i.e., when there is no spike-in splitting). For marks/antibodies not in this list the total mapped reads after flT3 will be used for normalization factor calculation.
type: string
default: H3K9me3,H3K27me3
Whether to save the intermediate files from the coverage calculation step. These include the coverage files in bedGraph format.
type: boolean
Minimum number of total mapped reads required to include a BAM in any normalization. For example, this prevents having an empty exogenous input control BAM when no reads map to the spike-in genome.
type: integer
default: 1
Whether to skip the generation of average bigWig files across replicates for each sample group.
type: boolean
Whether to skip the generation of exogenous bigWig files for each sample.
type: boolean
default: true
Whether to skip the generation of a table with the bin value for all the samples.
type: boolean
Whether to skip correlation of multibigwigsummary output.
type: boolean
default: true
Whether to skip PCA of multibigwigsummary output.
type: boolean
default: true

Options to adjust downsampling criteria.

Method to downsample BAM files. Leave empty to disable downsampling.
type: string
options: min_endo_across, min_exo_across, min_endo_by_type, min_exo_by_type
Help text

The min_endo_across method will downsample all IPs and inputs of the same antibody to the minimum number of endogenous reads across all IPs and inputs of the same antibody. The min_exo_across method will downsample all IPs and inputs of the same antibody to the minimum number of exogenous reads across all IPs and inputs of the same antibody. The min_endo_by_type method will downsample all IPs to the minimum number of endogenous reads across all IPs of the same antibody, and all the inputs to the minimum number of endogenous reads across all inputs of the same antibody. The min_exo_by_type method will downsample all IPs to the minimum number of exogenous reads across all IPs of the same antibody, and all the inputs to the minimum number of exogenous reads across all inputs of the same antibody.

Threshold of endogenous mapped reads for downsampling. For downsampling methods that search for the minimum endogenous mapped reads across a set of samples, this parameter will determine the lower limit of that minimum. In other words, the minimum mapped endogenous reads should be the minimum number above the downsampling_endo_threshold. If there are no totals above this threshold, then the (true) minimum is used.
type: integer
default: 10000000
Threshold of exogenous mapped reads for downsampling. For downsampling methods that search for the minimum exogenous mapped reads across a set of samples, this parameter will determine the lower limit of that minimum. In other words, the minimum mapped exogenous reads should be the minimum number above the downsampling_exogenous_threshold. If there are no totals above this threshold, then the (true) minimum is used.
type: integer
default: 300000
Strategy to downsample BAM files. See Picard for more details.
type: string
options: ConstantMemory, HighAccuracy, Chained
default: Chained
Accuracy of downsampling BAM files. See Picard for more details.
type: number
default: 0.000001
Save the intermediate BAM files from the downsampling step.
type: boolean
default: true

Options to adjust transposable element (TE) counting criteria.

Skip transposable element (TE) counting.
type: boolean
default: true
Skip local TE counting.
type: boolean
default: true
Skip gzip compression of TEcount output files.
type: boolean
Skip gzip compression of TElocal output files.
type: boolean
Method to use for TE counting. See TEcount's documentation for more details.
type: string
options: multi, uniq
default: multi
Path to a different gene GTF file for counting with TEcount and TElocal. If not provided, the main --gtf file will be used to generate a gene index suitable for TEcount or TElocal, unless the respective prebuilt gene index files are provided.
type: string
Path to the prebuilt gene index file (.ind) for counting with TEcount. If not provided, this file in --gtf will be used to generate a gene index suitable for TEcount.
type: string
Path to the prebuilt gene index file (.ind) for counting with TElocal. If not provided, this file in --gtf will be used to generate a gene index suitable for TElocal.
type: string
Path to the TE GTF file for counting with TEcount or TElocal. If not provided, this file will be used to generate a TE index suitable for TEcount or TElocal.
type: string
Path to the prebuilt TE index file (.ind) for counting with TEcount. If not provided, this file in --te_gtf will be used to generate a TE index suitable for TEcount.
type: string
Path to the prebuilt TE index file (.locInd) for local (instance) TE counting with TElocal. If not provided, this file in --te_gtf will be used to generate a TE index suitable for TElocal.
type: string

Options to adjust peak calling criteria.

Skip peak calling altogether, for every peak caller listed in peak_callers.
type: boolean
Comma-separated list of peak calling algorithms to use.
type: string
default: macs3
pattern: ^(macs3|genrich|consenrich|epic2|mace|dpeak|dpos|dregion|dtriple|edd|denopa|seacr)(,(macs3|genrich|consenrich|epic2|mace|dpeak|dpos|dregion|dtriple|edd|denopa|seacr))*$
Help text

One or more of macs3, genrich, consenrich, epic2, mace, dpeak, dpos, dregion, dtriple, edd, denopa, seacr, given as a comma-separated list without spaces, e.g. --peak_callers macs3,genrich. Every listed peak caller is run on all applicable samples. Use --skip_peak_calling to switch peak calling off entirely. denopa is currently not supported: no usable container for it exists yet.

Whether to call narrow peaks with MACS3 and Genrich. Default is to call broad peaks.
type: boolean
Type of statistical cutoff to use for peak calling.
type: string
options: qvalue, pvalue
default: qvalue
Specifies narrow cutoff value (p-value or q-value) for MACS3 and Genrich peak calling.
type: number
default: 0.01
Specifies broad cutoff value (p-value or q-value) for MACS3 and Genrich peak calling.
type: number
default: 0.1
Number of biological replicates required from a given condition for a peak to contribute to a consensus peak.
type: number
default: 1
Help text

If you are confident you have good reproducibility amongst your replicates then you can increase the value of this parameter to create a 'reproducible' set of consensus peaks. For example, a value of 2 will mean peaks that have been called in at least 2 replicates will contribute to the consensus set of peaks, and as such peaks that are unique to a given replicate will be discarded. A value of -1 means that a peak must have been called in all replicates to contribute to the consensus set of peaks.

Maximum distance between peaks allowed for peaks to be merged for consensus peak definition. --consensus_peaks_max_dist 0 means overlapping and/or book-ended features are merged.
type: integer
default: 0
Used to extend reads in 5'->3' direction to fix-sized fragments when processing ATAC-seq data, for which MACS3 bypasses building the shifting model (--nomodel). See MACS3 documentation for more details.
type: integer
default: 250
Size of the small local region around peak regions to calculate the maximum lambda as local lambda.
type: integer
default: 1500
Help text

By default, MACS3 considers 1000 bp for small local region (--slocal). You can tweak this according to your project. Remember that if the region is set too small, a sharp spike in the input data may kill a significant peak. See MACS3 documentation for more details.

Size of the large local region around peak regions to calculate the maximum lambda as local lambda.
type: integer
default: 10000
Help text

By default, MACS3 considers 10000 bp for large local region (--llocal) which captures the bias from a long-range effect like an open chromatin domain. You can tweak this according to your project. See MACS3 documentation for more details.

Instruct MACS3 to create bedGraph files normalised to signal per million reads.
type: boolean
default: true
Skip MACS3 peak QC plot generation.
type: boolean
Skip annotation of MACS3 and consensus peaks with HOMER.
type: boolean
Skip consensus peak generation, annotation and counting.
type: boolean
Skip generation of the consensus peak profile plot.
type: boolean
Mode to use for the consensus peak profile plot. See the deepTools documentation for more details.
type: string
options: reference_point, scale_regions
default: reference_point
Reference point to use for the consensus peak profile plot when --consensus_plotprofile_mode is set to reference_point. See the deepTools documentation for more details.
type: string
options: TSS, TES, center
default: center
Length of the region body to plot for the consensus peak profile plot when --consensus_plotprofile_mode is set to scale_regions. See the deepTools documentation for more details.
type: integer
default: 1000
Length of the upstream region to plot for the consensus peak profile plot. See the deepTools documentation for more details.
type: integer
default: 6000
Length of the downstream region to plot for the consensus peak profile plot. See the deepTools documentation for more details.
type: integer
default: 6000
Whether to skip regions with zero coverage in the consensus peak profile plot. See the deepTools documentation for more details.
type: boolean
default: true
Whether to plot all samples in a group together in the consensus peak profile plot. See the deepTools documentation for more details.
type: boolean
default: true
Whether to include the input CISRPM signal in the gene or consensus peak profile plots.
type: boolean
Skip bdgcmp calculation with MACS3.
type: boolean
default: true
Save intermediate bedGraph/bigWig files generated by the MACS3 bdgcmp step.
type: boolean
Bin size (Kb) for EDD peak calling. Autoestimated if left blank. See EDD documentation for more details.
type: string
Gap penalty for EDD peak calling. Autoestimated by EDD if left blank. See EDD documentation for more details.
type: string
Number of Monte Carlo trials for EDD peak calling. See EDD documentation for more details.
type: integer
default: 10000
FDR threshold for EDD peak calling. See EDD documentation for more details.
type: number
default: 0.01
Genrich narrow gap parameter.
type: integer
default: 100
Genrich broad gap parameter.
type: integer
default: 200
Instruct Genrich to save pileup files.
type: boolean
default: true
Instruct Genrich to save p-values.
type: boolean
default: true
Instruct Genrich to save intervals.
type: boolean
default: true
Size of the windows to scan the genome in epic2. Bin size is the smallest possible island.
type: integer
default: 250
In epic2 peak calling, remove all islands with an FDR above this cutoff.
type: number
default: 0.01
Skip peak calling with the ENCODE ChIP-seq pipeline.
type: boolean
default: true
For each sample, if the ratio of depth between input controls is higher than this, then a pooled input control will be used for all sample replicates.
type: number
default: 1.2
Set the IDR value for a group of multi summit peaks (a group of peaks with the same chr/start/stop but different summits) to the best value across all of these peaks. This is a work around for peak callers that don't do a good job splitting scores across multi summit peaks (e.g. MACS). If set in conjunction with --plot two plots will be created - one with alternate summits and one without. Use this option with care. See the IDR documentation for more details.
type: boolean
default: true
Report statistics for peaks with a global IDR below this value but return all peaks
type: number
default: 0.05
Filter peaks with an IDR above this value.
type: number
default: 0.05
Threshold on the number of peaks to call with SPP for the ENCODE-based processing.
type: integer
default: 300000
Set peaks with a score above this value to this value.
type: number
default: 1000
Minimum fragment length to exclude when calculating cross-correlation metrics for the ENCODE-based processing.
type: integer
default: -500
Maximum fragment length to exclude when calculating cross-correlation metrics for the ENCODE-based processing. If not provided max(params.read_length + 10, 100) will be used.
type: integer
SEACR specifies returns the top n fraction (between 0 and 1) of peaks based on total signal within peaks. This is only used if there are no controls included with the samples and if --use_control is false
type: number
default: 0.05
SEACR normalization.
type: string
options: non, norm
default: non
SEACR stringency.
type: string
options: stringent, relaxed
default: stringent
Set the k-mer size for MACE peak calling. Kmer size [6,12] to correct nucleotide composition bias. kmerSize < 0.5*read_length. Larger KmerSize might make program slower. Set kmerSize = 0 to turn off nucleotide compsition bias correction. default=6. See the MACE documentation for more details.
type: integer
Chromosome chunk size. Each chomosome will be cut into small chunks of this size. Decrease chunk size will save more RAM. default=100000 (bp). See the MACE documentation for more details.
type: integer
Reference reads count (default = 10 million). Sequencing depth will be normailzed to this number, so that wig files are comparable between replicates. See the MACE documentation for more details.
type: integer
phred scaled mapping quality threshhold to determine “uniqueness” of alignments. default=30. See the MACE documentation for more details.
type: number
Method used to consolidate replicates and reduce noise. “EM” = Entropy weighted mean, “AM”=Arithmetic mean, “GM”=Geometric mean, “SNR”=Signal-to-noise ratio. See the MACE documentation for more details.
type: string
options: EM, AM, GM, SNR
Pvalue cutoff for border detection and subsequent border pairing. default=0.05. See the MACE documentation for more details.
type: number
Maximum distance allowed for border pairing. default=100 (bp). See the MACE documentation for more details.
type: integer
Peaks (i.e. borders) located closely within this window will be merged. default=5 (bp). See the MACE documentation for more details.
type: integer
Background window size used to determine background signal level. default=100 (bp). See the MACE documentation for more details.
type: integer
Minimum coverage signal used to build model (i.e. estimate optimal peak pair size). default=2.0. See the MACE documentation for more details.
type: number
Whether to save the intermediate files from the MACE peak calling step. These include the MACE peak files in bedGraph format.
type: boolean

Options to skip various steps within the workflow.

Skip FastQC.
type: boolean
Path to a tab-delimited file containing the spike-in barcode targets, ids, and sequences. See the sc_spikein_barcodes.tsv for an example. This is only useful for CUT&Tag or CUT&RUN experiments with SNAP-CUTANA™ Spike-ins (See https://support.epicypher.com/v1/docs/analyzing-snap-cutana-spike-in-controls-and-expected-results).
type: string
pattern: ^\S+\.tsv(\.gz)?$
Whether to skip the extraction of spike-in barcodes from FASTQ files. This is useful for CUT&Tag or CUT&RUN experiments with SNAP-CUTANA™ Spike-ins (See https://support.epicypher.com/v1/docs/analyzing-snap-cutana-spike-in-controls-and-expected-results).
type: boolean
Skip Picard CollectMultipleMetrics.
type: boolean
Skip Preseq.
type: boolean
default: true
Use vst transformation instead of rlog with DESeq2.
type: boolean
default: true
Help text
Skip deepTools plotProfile.
type: boolean
Mode to compute the matrix for deepTools plotProfile. See the deepTools documentation for more details.
type: string
options: scale_regions, reference_point
default: scale_regions
Reference point for deepTools plotProfile when --genes_plotprofile_mode is reference_point. See the deepTools documentation for more details.
type: string
options: TSS, TES, center
default: TSS
Region body length (in bp) for deepTools plotProfile when --genes_plotprofile_mode is scale_regions. See the deepTools documentation for more details.
type: integer
default: 1000
Upstream distance (in bp) from the reference point for deepTools plotProfile when --genes_plotprofile_mode is reference_point. See the deepTools documentation for more details.
type: integer
default: 2000
Downstream distance (in bp) from the reference point for deepTools plotProfile when --genes_plotprofile_mode is reference_point. See the deepTools documentation for more details.
type: integer
default: 2000
Whether to skip regions with zero coverage in all samples when computing the matrix for deepTools plotProfile. This is determined BEFORE any applicable pseudocount is added. See the deepTools documentation for more details.
type: boolean
Skip deepTools plotFingerprint.
type: boolean
Skip Phantompeakqualtools QC metrics and plots.
type: boolean
Skip DESeq2 PCA and heatmap plotting.
type: boolean
Skip IGV.
type: boolean
Skip MultiQC.
type: boolean
Skip all QC steps except for MultiQC.
type: boolean

Options to adjust SCAR-seq and OK-seq analysis criteria.

For single-end SCAR-seq or OK-seq data, the fragment size (in bp) to extend reads when calculating coverage for partition/RFD analysis. For paired-end data, coverage of intervals is calculated from the left point of a read pair to the right point.
type: integer
default: 250
Window/bin size (in bp) for dividing chromosomes when calculating coverage for partition/RFD analysis in SCAR-seq or OK-seq experiments. See this reference for more details.
type: integer
default: 1000
Step size (in bp) for sliding window when calculating coverage for partition/RFD analysis in SCAR-seq or OK-seq experiments. Default is the same as --partition_window_size to create fixed-size book-ended bins instead of sliding windows.
type: integer
default: 1000
Smoothing radius for partition/RFD signal smoothing. Smoothing is computed across consecutive book-ended bins, performing a uniform blur considering the neighboring bins on each side. See this reference for more details.
type: integer
default: 30
Radius for nonparametric derivative estimation of RFD signal to detect sharp transitions between neighboring bins for initiation zone detection. See this reference for more details.
type: integer
default: 30
Radius for zero-crossing detection when defining replication initiation zones. See this reference for more details.
type: integer
default: 1
RPM (reads per million) cutoff for bins to be included in downstream partition analysis. Bins with normalized read-counts above this threshold will be included. See this reference for more details.
type: number
default: 0.3
Distance (kb) up- and downstream of each initiation zone to plot.
type: integer
default: 100
Help text

Independent of partition_iz_rm_overlap_range: when partition_plot_range is the larger of the two, the plotted flanks can reach neighbouring initiation zones and a partition bin may then contribute to more than one profile.

Distance (kb) around each initiation zone used to discard overlapping ones.
type: integer
default: 100
Help text

Any initiation zone whose window of this radius overlaps another one is discarded before plotting. Set to 0 to skip the removal of overlapping initiation zones entirely and keep them all. Independent of partition_plot_range, so the plotted range can be widened without discarding more initiation zones.

Whether to exclude scaffolds from partition plots. Scaffolds are defined as chromosomes whose name begins with 'chrUn' or contains a dot ('.').
type: boolean
default: true
Comma-separated list of chromosomes to exclude from partition analysis. Typically sex chromosomes and mitochondrial chromosomes are excluded. See this reference for more details.
type: string
default: chrX,chrY,chrM
Whether to skip the grouped partition plots, which combine all samples of the same antibody in a single plot. Per-sample partition plots are still generated.
type: boolean
Whether to save the intermediate files from the partition/RFD analysis step.
type: boolean

Options to adjust Repli-seq replication-timing (RT) track analysis.

Bin size (in bp) used by deepTools multiBamSummary when counting reads per window for Repli-seq replication-timing (RT) tracks (e.g. 5000, 10000, 20000, 50000, 100000 for 5/10/20/50/100 kb resolution).
type: integer
default: 50000
How to combine early/late biological replicates when calculating the Repli-seq RT ratio. paired computes the ratio per matched early/late biological-replicate pair (matched by biological_replicate) and averages; pooled averages replicates within each phase first, then computes one ratio; auto chooses between the two via a correlation heuristic (see --repliseq_pairing_threshold).
type: string
options: auto, paired, pooled
default: pooled
cpm only CPM-normalizes each Repli-seq replicate; qn additionally quantile-normalizes between replicates, on top of CPM.
type: string
options: cpm, qn
default: qn
Smoothing strategy applied to the raw Repli-seq RT track.
type: string
options: loess, roll, none
default: loess
Where smoothing enters the Repli-seq RT calculation. replicate smooths each replicate's normalized coverage track and then takes the early/late ratio; ratio takes the ratio first and smooths that. Only affects the smoothed track; the raw track is always the unsmoothed ratio.
type: string
options: replicate, ratio
default: replicate
Floor smoothed replicate coverage at zero before taking the Repli-seq early/late ratio. Coverage cannot be negative, but a loess fit can undershoot below it. Only applies with --repliseq_smooth_stage replicate.
type: boolean
default: true
Orientation of the Repli-seq log2 ratio. early_over_late makes high values early-replicating, which is the usual convention; late_over_early inverts it.
type: string
options: early_over_late, late_over_early
default: early_over_late
Degree of the local polynomial fitted by loess when smoothing Repli-seq RT tracks.
type: integer
options: 1, 2
default: 2
Loess fitting family used when smoothing Repli-seq RT tracks. symmetric reweights to resist outliers, but is undefined on sequences where over half the residuals are exactly zero, in which case it falls back to gaussian for that sequence.
type: string
options: gaussian, symmetric
default: gaussian
Width (in bp) of the loess smoothing window used when --repliseq_smooth loess. It is converted to a per-sequence span, so the same genomic scale is smoothed over on every sequence regardless of its length.
type: integer
default: 500000
Loess span given directly, as a fraction of the bins on each sequence, used when --repliseq_smooth loess. Overrides --repliseq_loess_window when set. Because it is a fraction rather than a fixed width, a long chromosome is smoothed over a proportionally wider stretch of sequence than a short one.
type: number
Rolling-mean window size, in bins, used when --repliseq_smooth roll.
type: integer
default: 5
Correlation method used by the --repliseq_replicate_method auto heuristic.
type: string
options: spearman, pearson
default: spearman
Under --repliseq_replicate_method auto, paired is chosen when (matched-replicate-pair average correlation - mismatched-replicate-pair average correlation) is at least this value; otherwise pooled is used.
type: number
default: 0.02
Exclude scaffolds and alternate sequences from the Repli-seq RT track. deepTools multiBamSummary bins every sequence in the BAM header, and these are short and usually barely covered, which makes their RT values meaningless. A sequence counts as one when its name begins with chrUn, ends with _random, _alt or _fix, or contains a dot. They are dropped before CPM normalization, so the normalization reflects primary-chromosome depth only.
type: boolean
default: true
Fewest reads a bin needs across the early and late fractions combined to appear in the .covered copy of the Repli-seq ratio track. A bin with no reads in either fraction still gets a ratio of exactly log2(1/1) = 0, which is indistinguishable from a genuinely mid-replicating bin, so replication-timing domains are called on the covered copy while the unfiltered track keeps every bin.
type: integer
default: 1
Skip calling domains of constant replication timing from the Repli-seq ratio track.
type: boolean
Which Repli-seq ratio track to segment into replication-timing domains. The default both segments the raw and the smoothed track. Set smooth or raw to segment only one.
type: string
options: smooth, raw, both
default: both
How to call each replication-timing domain. binary calls every domain early or late from the sign of its segment mean; three_way additionally calls domains whose mean sits within --repliseq_domain_threshold of zero as intermediate, i.e. replicating throughout S phase.
type: string
options: three_way, binary
default: three_way
Half-width of the intermediate band around a segment mean of zero, in log2 ratio units. Only used with --repliseq_domain_classification three_way.
type: number
default: 0.1
Significance level for accepting a change point in DNAcopy's circular binary segmentation of the Repli-seq ratio track.
type: number
default: 0.01
Smallest number of bins a replication-timing domain may contain. DNAcopy's segment() accepts 2 to 5.
type: integer
options: 2, 3, 4, 5
default: 2
Merge adjacent replication-timing domains whose segment means differ by fewer than this many standard deviations. Zero or less turns the merging off.
type: number
default: 1
Run DNAcopy's smooth.CNA() before segmenting, which pulls single-bin outliers towards their neighbours so that one noisy bin does not become its own domain.
type: boolean
default: true
Random seed for replication-timing domain calling. DNAcopy locates change points with a permutation test, so without a fixed seed the same track segments slightly differently on every run.
type: integer
default: 1
Skip classifying genes by the S-phase fraction with the highest read density over their gene body.
type: boolean
Fewest reads a gene needs summed over every S-phase fraction before it is classified. Genes below it are reported as unclassified, because with only a handful of reads the fraction with the highest density is decided by sampling noise.
type: integer
default: 20
Fewest CPM by which the winning S-phase fraction must exceed the runner-up for a gene to be classified. Zero accepts any winner.
type: number
default: 0
Skip the diagnostic plots of the Repli-seq gene classification. The classification table and BEDs are still written.
type: boolean
Skip the diagnostic plots of the Repli-seq replication-timing domains. The domain BEDs and QC are still written.
type: boolean
Whether to save the intermediate files from the Repli-seq RT-track analysis step (raw counts tables and raw, unsmoothed bedGraph tracks).
type: boolean

Options to adjust high-resolution (16-fraction) Repli-seq analysis.

Bin size (in bp) used by deepTools multiBamSummary when counting reads per window for high-resolution Repli-seq. 50 kb is the usual resolution: a 30 min BrdU pulse at a fork speed of 1.8 kb/min labels at least 50 kb per fork.
type: integer
default: 50000
How each S phase fraction is normalized before the Repli-seq array is built. g1_filter drops the bins a G1 whole-genome control does not cover, the mappability correction, and keeps the fractions as reads per million. g1_ratio additionally replaces each fraction with log2 of its ratio to the control, floored at zero: a stricter correction, but because every fraction and the control share the same mean the floor zeroes about half of each fraction, and on H1 it collapses the initiation-zone calls from ~2950 to 9, so it should not be used to call features. rpm corrects for sequencing depth only. auto uses g1_filter when a G1 fraction is present in the samplesheet and rpm otherwise.
type: string
options: auto, none, rpm, g1_filter, g1_ratio
default: auto
Value of the rt_fraction samplesheet column that marks the G1 whole-genome-sequencing control.
hidden
type: string
default: G1
Bins whose G1 control coverage is at or below this many reads per million are dropped, since their ratio to the control is undefined.
type: number
default: 0
Comma-separated sequences to drop before building the array, matched by name. The default removes the sex chromosomes, whose copy number differs from the autosomes, and the mitochondrial genome, which does not replicate on the nuclear schedule. Set to an empty string to keep every sequence.
type: string
default: chrX,chrY,chrM
Comma-separated list of sequences to restrict the array to (e.g. chr1,chr2,chr3), which also sets the order of the output. Leave unset to use every sequence that survives the scaffold and sex-chromosome filtering.
type: string
BIRCH clustering threshold used to group bins into replication features. Lower values split the array into more, smaller clusters.
type: number
default: 0.5
Number of BIRCH clusters to reduce the subclusters to. Leave unset to keep the subclusters themselves.
hidden
type: integer
Valleys in the replication-timing profile up to this size (in bp) are called termination sites; larger ones are called late constant-timing regions.
type: integer
default: 100000
Order in which overlapping feature calls are resolved into the partition, in which every analysed bin belongs to exactly one feature. Later entries win, so the last one listed takes precedence.
type: string
default: TTR,breakage,CTR,termination,IZ
Assumed length of S phase in hours. A timing transition region is traversed by a single fork, so its length divided by the stretch of S phase it spans gives that fork's speed; this sets how long that stretch is in minutes. The method's paper assumes 10 h, the longest of the five cell lines it profiles, and reports a median of 1.7-2.0 kb/min.
type: number
default: 10
Comma-separated list of chromosomes to draw a Repli-seq heatmap page for. Leave unset to draw every chromosome in the array.
type: string
Chromosome whose Repli-seq heatmap is shown in the MultiQC report. Defaults to the first one drawn.
type: string
How many pages of example features to add to the plots PDF. Each page draws one randomly chosen instance of every feature shape: the cluster-centroid rank profile with the feature's bins picked out, over the heatmap of the same bins. Set to 0 to leave them out.
type: integer
default: 5
How many bins to show either side of an example feature. Smaller zooms in on the shape itself, larger gives it more surrounding context.
type: integer
default: 4
Seed for choosing which feature instances the example pages show, so that a rerun draws the same ones.
hidden
type: integer
default: 1
Skip the high-resolution Repli-seq heatmaps and feature summary plots.
type: boolean

Parameters used to describe centralised config profiles. These should not be edited.

Help text

The centralised nf-core configuration profiles use a handful of pipeline parameters to describe themselves. This information is then printed to the Nextflow log when you run a pipeline. You should not need to change these values when you run a pipeline.

Git commit id for Institutional configs.
hidden
type: string
default: master
Base directory for Institutional configs.
hidden
type: string
default: https://raw.githubusercontent.com/nf-core/configs/master
Help text

If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.

Institutional config name.
hidden
type: string
Institutional config description.
hidden
type: string
Institutional config contact information.
hidden
type: string
Institutional config URL link.
hidden
type: string

Less common options for the pipeline, typically set in a config file.

Help text

These options are common to all nf-core pipelines and allow you to customise some of the core preferences for how the pipeline runs.

Typically these options would be set in a Nextflow config file loaded for all pipeline runs, such as ~/.nextflow/config.

Display help text.
hidden
type: boolean
Method used to save pipeline results to output directory.
hidden
type: string
options: symlink, rellink, link, copy, copyNoFollow, move
default: copy
Help text

The Nextflow publishDir option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See Nextflow docs for details.

Number of genomic bins to use when calculating deepTools fingerprint plot.
hidden
type: integer
default: 500000
Email address for completion summary, only when pipeline fails.
hidden
type: string
pattern: ^([a-zA-Z0-9_\-\.]+)@([a-zA-Z0-9_\-\.]+)\.([a-zA-Z]{2,5})$
Help text

An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.

Send plain-text email instead of HTML.
hidden
type: boolean
File size limit when attaching MultiQC reports to summary emails.
hidden
type: string
default: 25.MB
pattern: ^\d+(\.\d+)?\.?\s*(K|M|G|T)?B$
Do not use coloured log outputs.
hidden
type: boolean
Custom config file to supply to MultiQC.
hidden
type: string
Boolean whether to validate parameters against the schema at runtime
hidden
type: boolean
default: true
Display the full detailed help message.
type: boolean
Display hidden parameters in the help message (only works when --help or --help_full are provided).
type: boolean
Display version and exit.
hidden
type: boolean
Custom MultiQC yaml file containing HTML including a methods description.
type: string
Base URL or local path to location of pipeline test dataset files
hidden
type: string
default: https://raw.githubusercontent.com/nf-core/test-datasets/
Base URL or local path to location of grothlab/crepas-specific test dataset files.
hidden
type: string
Suffix to add to the trace report filename. Default is the date and time in the format yyyy-MM-dd_HH-mm-ss.
hidden
type: string